EP 0754764 B1 20030409 - Method and reagent for the specific determination of mRNA
Title (en)
Method and reagent for the specific determination of mRNA
Title (de)
Verfahren und Reagenz zur spezifischen Bestimmung von mRNA
Title (fr)
Procédé et réactif pour la détermination spécifique d'ARNm
Publication
Application
Priority
DE 19526431 A 19950721
Abstract (en)
[origin: EP0754764A2] Specific determn. of nucleic acid (I) contg. poly-A sequences (II) comprises: (a) treating sample material with a lysis or hybridisation buffer and opt. homogenising it; (b) treating the sample soln. with a labelled poly- or oligo-nucleotide probe (III) or nucleotide deriv. with ≥ 1 sequence complementary to (II) or (I); (c) transfer of an aliquot of the mixt. to a heat-stable, RNase-free reaction vessel coated with an organic cpd. (A) and incubating for ≥ 10 secs. at 4-50 degrees C; (d) removing supernatant from the vessel and washing several times with buffer; (e) adding to the vessel an enzyme with reverse transcriptase (RT) activity and opt. heat-stable DNA polymerase activity, and incubating at 30-75 degrees C; (f) opt. removing the reaction mixt. and washing with buffer; (g) opt. adding a mixt. of heat-stable DNA polymerase and 2 specific primers, then incubation under amplification conditions, and (h) opt. recovering the DNA, esp. by electrophoresis, and detecting it. Also new are kits for this process.
IPC 1-7
IPC 8 full level
G01N 33/566 (2006.01); C07H 21/02 (2006.01); C07H 21/04 (2006.01); C12N 15/09 (2006.01); C12P 19/34 (2006.01); C12Q 1/48 (2006.01); C12Q 1/68 (2006.01); C12Q 1/6806 (2018.01); C12R 1/91 (2006.01)
CPC (source: EP US)
C12Q 1/6806 (2013.01 - EP US)
Designated contracting state (EPC)
AT CH DE ES FR GB IT LI NL SE
DOCDB simple family (publication)
EP 0754764 A2 19970122; EP 0754764 A3 19990421; EP 0754764 B1 20030409; AT E236995 T1 20030415; CA 2181417 A1 19970122; CA 2181417 C 20090922; DE 19526431 A1 19970123; DE 59610324 D1 20030515; ES 2193215 T3 20031101; JP H0928398 A 19970204; US 5955271 A 19990921
DOCDB simple family (application)
EP 96111586 A 19960718; AT 96111586 T 19960718; CA 2181417 A 19960717; DE 19526431 A 19950721; DE 59610324 T 19960718; ES 96111586 T 19960718; JP 20914496 A 19960718; US 68101596 A 19960722